Laboratory Experiment

Cite This: J. Chem. Educ. XXXX, XXX, XXX−XXX pubs.acs.org/jchemeduc

Chemical Curiosity on Campus: An Undergraduate Project on the

Structure and Wettability of Natural Surfaces
Anthony Katselas,† Alice Motion,† Chiara O’Reilly,‡ and Chiara Neto*,†
†
School of Chemistry and Sydney Nano Institute, The University of Sydney, Sydney, New South Wales 2006, Australia
‡
Department of Art History, The University of Sydney, Sydney, New South Wales 2006, Australia
*
S Supporting Information

ABSTRACT: An experiment which investigates the wettability of natural

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surfaces was developed for undergraduate students, which allows them to

explore concepts of surface structure, wettability, superhydrophobic
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surfaces, and self-cleaning. The students choose their own samples of

leaves and flowers present in their local environment, examine their surface
structure and chemistry by microscopy and contact angle measurements,
and, using their understanding of the literature on the wettability of rough
surfaces, relate the two. The experiment is student-focused in that it gives
students agency in choosing the samples to study, increases their
confidence, and develops curiosity in their local environment.

KEYWORDS: Upper-Division Undergraduate, Inquiry-Based/Discovery Learning, Interdisciplinary/Multidisciplinary,

Physical Chemistry, Materials Science, Surface Science, Hands-On Learning/Manipulatives

P lant and animal species have evolved over centuries to

survive in harsh yet bountiful conditions. Plant species
such as the leaf of the sacred lotus have long been known to
scientists for their extreme water repellency and self-cleaning
properties, which provide a functional basis for survival.1−4
These properties are made possible by the intrinsic hydro-
phobicity of epicuticular wax crystals which adorn the plant’s
leaf surface.5 On their own, waxy hydrocarbon-based coatings
provide a chemical basis of moderate water repellency.6 Yet, it
is the micro- and nanoscale roughness of these waxy crystal
structures that achieves the property of superhydrophobicity.1,7
Superhydrophobicity is one of a number of coveted surface
properties found in nature. Antibiofouling ability, antibacterial
properties, self-healing ability, structural coloration, and dry
adhesion are other well-known properties of interest.8−11 The
field of biomimetics focuses on studying biological systems
displaying these properties, taking inspiration, and using the
properties to solve real world problems.2 Biomimetics is an
interdisciplinary field, which brings together engineering,
chemistry, and biology, making it an interesting topic for
many students. However, biomimetics is seldom taught as part
of undergraduate science degree programs. Surface chemistry
provides the basic characterization tools used in biomimetic
research and offers the benefit of introducing biological system
characterization as a final undergraduate year research project.
The undergraduate research project described here has
inspired students into pursuing further research studies and
offers academic staff the opportunity to explore new research
directions.
© XXXX American Chemical Society and
Division of Chemical Education, Inc.
Characterizing a surface’s wettability is an important aspect
of biomimetic research. Wettability provides qualitative
information on the chemistry of a material’s surface, since
the degree of wetting is dependent on the intermolecular
interactions between the surface and a liquid droplet.12 When
water droplets only partially wet the surface, the surface
energies of the relevant liquid−vapor, liquid−solid, and solid−
vapor phases achieve a balance, resulting in a contact angle
measured at the three-phase contact line (solid/liquid/
vapor),12 as shown in Figure 1.
The equilibrium contact angle θ shown in Figure 1 is defined
by Young’s equation:
Figure 1. (a) Schematic of a liquid droplet on a solid substrate, with
the equilibrium contact angle θ shown, taken at the point where the
liquid/vapor contact line meets the solid interface. (b) A droplet
suspended in the Cassie−Baxter state, in which the space between
surface features contains air pockets. (c) A droplet in a collapsed
Wenzel state partially penetrates the surface roughness, replacing the
air pockets with water, resulting in droplet pinning.
Received: April 3, 2019
Revised: June 3, 2019
A DOI: 10.1021/acs.jchemed.9b00324
J. Chem. Educ. XXXX, XXX, XXX−XXX
Journal of Chemical Education
γsv − γsl
cos θ =
γlv (1)
where γsv, γsl, and γlv represent the surface energy of the solid−
vapor, solid−liquid, and liquid−vapor interfaces, respectively.12
As real surfaces have heterogeneity and roughness, in
practice multiple contact angles are measured when the contact
line moves over the surface, and this is visualized by measuring
contact angle hysteresis, the difference between the advancing
and receding contact angle. The advancing contact angle, θA, is
measured by increasing the water droplet volume and
measuring the maximum contact angle before the contact
line moves outward; similarly, the receding contact, θR, is
measured by decreasing the droplet volume and measuring the
minimum contact angle before the contact line moves
inward.12 Atomically smooth hydrophobic surfaces typically
display a low hysteresis of around 10−15°,13,14 with more
heterogeneous surfaces displaying higher hysteresis values.2,12
The effect of surface roughness is generally to amplify the
wetting character of a surface and can be explained (at least for
large droplets) by the Wenzel and Cassie−Baxter models,
depicted in Figure 1.2,15,16 In the Wenzel wetting state, a water
droplet may show a very high contact angle (about 150°), but
as the water partially penetrates the roughness of the surface,
high droplet pinning and, therefore, higher contact angle
hysteresis (>20°) result (Figure 1c).2 In the Cassie−Baxter
model the droplet is suspended over a mattress of air pockets
trapped within the roughness, and this limits droplet contact to
the top of roughness peaks. Functionally, this minimizes
droplet pinning, reduces contact angle hysteresis to a few
degrees, and aids droplet roll-off at low tilt angles.2
There are only a few papers that have introduced in the
chemical education literature the wettability of natural
surfaces,6,17 the concept of self-cleaning,18,19 and the relation-
ship between hysteresis and surface heterogeneity.19 Our
experiment is novel in that it brings these concepts together
across a broad range of natural plant species while also
incorporating microscopic inspection of samples. Our experi-
ment mimics more closely than previous works methods used
in biomimetic research,17,20−23 and it gives an opportunity for
inquiry-led learning, by guiding students to relate the micro-
and nanostructure of natural surfaces to their wetting
properties, and to deduce wetting states from their contact
angle hysteresis values.
Primarily the students work toward identifying whether
biological plant species chosen by them are naturally
superhydrophobic and, if so, if they also possess self-cleaning
properties. To demonstrate this understanding, the concepts
are applied in the laboratory using samples, leaves and flowers
of plants available in the local area, collected by students and
brought back to the laboratory for analysis using available
instrumentation. The plant samples were selected on the basis
of species available on campus at the University of Sydney or
accessible locally (Figure 2). In other countries, local species
could be used. Species that appear hairy or rough are most
likely to provide interesting microscopic structure that is easily
visible under the optical microscope, and these samples are
more likely to have interesting wetting properties.
Core skills in material science, optical and scanning electron
microscopy (where available), are developed through this
practical experiment. Further skills in surface characterization
using contact angle measurement are practiced, realizing the
functional implications of surface roughness on natural
B DOI: 10.1021/acs.jchemed.9b00324
Laboratory Experiment
Figure 2. Photographs of the flannel flower (a), jacaranda flower (b),
kangaroo paw flower (c), and the banksia cob (d), collected prior to
analysis.
surfaces, their superhydrophobicity, and their self-cleaning
properties. Students also gain an appreciation of cleanliness
when working in the field of surface chemistry. Seven
undergraduate students have tested the experiment as
individuals and in groups and repeated the contact angle
measurements multiple times. The full experiment takes
approximately 12 h to complete and was originally developed
as a summer research project to engage final year under-
graduates with higher research. Following its success, the
experiment was offered as an advanced first-year practical
module. This experiment is best performed individually over
four sessions as outlined in the Supporting Information.
■
EXPERIMENTAL OVERVIEW
Sample Preparation for Optical Microscopy and Contact
Angle Measurement
Detailed instructions are provided in Supporting Information.
The samples studied were the flowers of four species:
Anigozanthos f lavidus (kangaroo paw) and Banksia serrata
(banksia) flowers were collected from the Sydney University
campus, and the Actinotus helianthi (flannel flower) was
purchased from Sydney Wildflower Nursery, Heathcote, NSW,
Australia. The introduced Jacaranda mimosifolia (jacaranda)
was another species chosen for its historical association with
Sydney University and was also collected on campus. On
collection, surface contamination was minimized by using
gloves and tweezers, and samples were then stored in sealed
plastic vials. The samples were carefully dissected using
tweezers and scissors or a razor blade. The sample size for
contact angle measurement was decided on the basis of the
sample structure, ensuring the flattest and largest surface area
was available (several cm2, if possible). Students must pay
attention to reduce contamination of the samples; this includes
gloves for handling samples, containers, or tweezers, using
clean glassware and ultrapure water. Samples were then fixed to
a glass slide with double sided tape and pressed firmly down at
the edges to provide adequate adhesion. Samples were
characterized immediately after collection to gather results
based on the native state of the species.
Optical Microscopy
Samples were imaged using an optical microscope in reflection
mode (Nikon, Eclipse LV150) and viewed using a 5×
objective. Any image demonstrating the microscopic surface
structure was captured, and greater magnification objectives
were used where possible to magnify surface characteristics at
the microscopic level.
Contact Angle Goniometry
Contact angles were measured using a KSV Cam 200
goniometer. The static contact angle values were calculated
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Journal of Chemical Education
by fitting the water droplet baseline in the instrument software
utilizing the Young−Laplace model. Droplets of 10 μL (or
larger) of ultrapure Milli-Q water were used, containing <1
particulate/mL (of size >0.22 μm3) and having a resistivity of
18.2 MΩ cm. Advancing and receding contact angles were also
measured to determine the hysteresis by the mentioned
contact angle goniometer. If an equivalent instrument is not
available, a cell phone camera is sufficient to capture an image
of the water droplets, which is then analyzed using the freeware
ImageJ software to measure contact angles as outlined by
Lamour et al.24 A tilting table apparatus can also be easily
assembled from basic laboratory equipment as per the work of
Haines et al., which can replicate the results of advancing and
receding contact angle measurements.19
Samples Preparation for Scanning Electron Microscopy
Samples were prepared for scanning electron microscopy
(SEM) using two approaches. The first approach was by drying
overnight in a vacuum desiccator, which produces good
images, but the structures appeared dehydrated. The second
method was by a biological specimen fixation protocol which
can be used to achieve a more native appearance, but this
should be performed under the supervision of a laboratory
demonstrator. The dried samples were mounted on SEM stubs
using conductive carbon adhesive. Conductive silver paint was
used where greater contact was required, and then, the samples
were gold coated. Samples were imaged using a JEOL tabletop
SEM under high vacuum, aiming to capture the micro- or
nanostructures. Where an SEM is not available, a stronger
emphasis can be placed on the optical microscopy work. In this
case, students focus on the qualitative aspects of microscale
structures: at magnifications of ∼100× they may identify
features that contribute to roughness, such as trichomes, which
promote the superhydrophobic wetting states.
■
HAZARDS
During sample collection, students should wear gloves to
prevent any allergic irritation during contact with plant species.
The handling of glass slides and the use of razor blades should
be carried out with extreme caution; disposal should be in a
designated sharps receptacle.
■
RESULTS AND DISCUSSION
The four different flower species, jacaranda, kangaroo paw,
flannel flower, and banksia, were readily available at the time of
the experiment. The choice of samples allowed a comparative
study of the wettability properties of biological samples. After
slide preparation, the samples were analyzed using optical
microscopy first to observe the native surface morphology as
shown in Figure 3.
The flannel flower, kangaroo paw, and banksia flowers all
displayed similarities in their dense, hairy surface coverage with
hairy projections recognized as trichomes, visible on the micro-
and macroscales. The flannel flower displayed a relatively
uniform trichome length, with each primary structure
branched, and displaying a dense surface coverage (Figure
3a). The kangaroo paw displayed fewer but larger trichome
structures than the flannel flower, with numerous branches,
leading to much longer and rigid projections with greater
variation in trichome length (Figure 3c). The banksia flower
displayed waxy trichome hairs which at the surface were
heavily kinked. Much longer and straighter hairs were seen to
be layered over the top (Figure 3d). The jacaranda displayed
C DOI: 10.1021/acs.jchemed.9b00324
Laboratory Experiment
Figure 3. Optical micrographs displaying the native morphology of
the flannel flower (a), jacaranda flower (b), kangaroo paw flower (c),
and Banksia serrata flower (d). Image insets depict the contact angle
measurement on each sample.
trichomes, segmented into three portions. In Figure 3b the
attachment point of the trichomes to the flower epicuticle
appears dark blue, and the trichomes point downward,
becoming more transparent toward the tips. The vertical
alignment of the trichomes corresponds to the vertical
downward orientation of the flowers on the tree. The features
were oriented in the vertical plane (Figure 3b). The spacing
measured between surface features was regular, 109 ± 26 μm.
For each of the species, the surface was greasy to the touch
and, in the case of the banksia, left behind a waxy residue on
contact. This qualitative observation suggests that the surface
features may be made of a hydrocarbon-based material, which
is typical of epicuticular waxes.4,6,20
The second component of the experiment required students
to measure contact angles and contact angle hysteresis. Static
contact angles were measured on each plant species a total of
six times per species, across multiple samples, and an average
and standard deviation reported. The results are shown in
Table 1.
Table 1. Summary of Contact Angle and Contact Angle
Hysteresis Measurements with the Associated Standard
Deviation Values
Surface Contact Angle/deg Hysteresis/deg
Flannel flower 154 ± 1 25 ± 2
Jacaranda flower 157 ± 6 41 ± 5
Kangaroo paw flower 137 ± 13 n/a
Banksia flower 150 ± 8 22 ± 6
Considering the nature of epicuticular waxes, all samples
were not surprisingly hydrophobic with most samples
displaying contact angle values in the superhydrophobic
regime (θ > 150°). The kangaroo paw flower was the sample
with the lowest static contact angle value, and the value was
affected by large measurement variability, due to the irregular
surface structure. Overall, the scatter in the measurement of
static contact angle was high (except in the flannel flower) due
the irregular surface features. Both the flannel flower and
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banksia displayed comparable and high contact angle hysteresis
values, which suggest a rough surface chemistry and/or an
irregular surface. The high value of contact angle hysteresis
points to a Wenzel state,2,15,16 whereby the droplets make
direct contact with the full length of the surface features,
penetrating the roughness. This type of topography is unlikely
to trap air pockets between the features, and does not produce
an effective self-cleaning effect. The jacaranda flower displayed
the fewest number of surface projections and overall had flatter
topography, and it had the highest value of hysteresis of all
samples. Contact angle hysteresis could not be measured on
the kangaroo paw, due to the spontaneous collapse and
spreading of the water droplet within the porous flower
structure when increasing droplet volume.
For the SEM analysis component of the experiment, the
jacaranda flower was prepared using both the biological
fixation and vacuum drying methods for comparison (Figure
4). The fixed samples clearly displayed finger-like trichome
Figure 4. SEM micrographs of the jacaranda flower surface prepared
using the biological fixation protocol (a, b) and sample which was
vacuum-dried (c, d). The higher magnification images highlight the
nanoscale texture observed on the surface projections.
projections which were composed of 3−4 segments, with the
terminal segment drawing the feature into a point. The
projections ranged in length from 165 to 250 μm, with a width
from 33 to 46 μm. The projections also displayed secondary
ridge-like structures which were 2−10 μm in length and 0.7−
1.1 μm in width. Overall, the morphology of the fixed samples
appeared comparable to the native morphology (Figure 3b),
but the vacuum-dried sample of the jacaranda flower appeared
shriveled, relative to the structure observed in the optical
micrographs (Figure 3b), making it difficult to measure. This
observation highlights the importance of correct biological
D DOI: 10.1021/acs.jchemed.9b00324
Laboratory Experiment
sample fixation for SEM analysis and indirectly suggests that
the surface projections were heavily hydrated in their native
state.
If students do not have access to biological sample fixation,
conclusions may still be drawn from vacuum-dried samples, as
clear nanoscale surface features can be observed, and
inferences can be made from the correlation with optical
micrographs. SEM micrographs of the remaining samples
prepared through vacuum drying are shown in Figure 5. All of
Figure 5. SEM micrographs of the banksia serrata flower (a, b),
flannel flower (c, d), and kangaroo paw (e, f).
the samples display hair-like or branch-like projections. The
banksia flower (Figure 5a) displayed trichomes which were
more hair-like, whereas the flannel flower (Figure 5c)
trichomes appeared to branch from a central point into longer
branches, and the kangaroo paw (Figure 5e) appeared to
branch off from a central projection into numerous short
branches. At a higher magnification, the banksia flower (Figure
5b) and flannel flower (Figure 5d) trichomes displayed a
nanoscale wrinkled texture, unlike the kangaroo paw, which
appears smooth on the same scale (Figure 5f). It is not possible
to conclude that the nanoscale wrinkles observed are native
features and not a result of the vacuum drying process. What is
clear, however, is that the structures are much more robust
than those of the jacaranda flower, retaining their native
morphology at the microscale. It can be inferred that the
smoother surface of the kangaroo paw led to increased surface
wetting, and indeed, the kangaroo paw had the lowest water
contact angles out of all the species (Table 1).
In the lotus leaf, it is particularly the hierarchical (micro- and
nanoscale superimposed) roughness that is effective at trapping
air, achieving the Cassie−Baxter state. The self-cleaning ability
is related to the Cassie−Baxter state, because only in this
configuration does the topography allow capillary forces to
direct any dirt resting on the structure to adhere to the water
droplet.4 Self-cleaning species typically have an ultralow
hysteresis measurement of <10°.1,4,25 Given the high value of
hysteresis for all the studied species, none of them appear to be
clearly self-cleaning, but in a natural setting the natural
flexibility of the flower, and close to vertical orientation, might
help with the roll-off of droplets.
■
PEDAGOGICAL GOALS
In this experiment the concepts of wettability and surface
roughness and their methods of characterization are intro-
duced. Students investigate directly the properties of natural
surfaces and their applications in material science through
biomimicry. The skills practiced include the use of sample
preparation, measurements and interpretation of optical and
scanning electron micrographs, and contact angle measure-
J. Chem. Educ. XXXX, XXX, XXX−XXX
Journal of Chemical Education
ments. Studying samples on campus develops an individual and
collective curiosity for local plant species while improving the
skill of observation. These broader skills are essential to
scientific research and overall student confidence.
The observations by microscopy techniques inform on the
physical structure of the material and how such structures may
contribute toward a function. From the contact angle
measurements performed, students can deduce whether the
natural materials are superhydrophobic and/or self-cleaning
and better understand the underlying surface chemistry.
Students should demonstrate an understanding of the concepts
mentioned and have practiced the skills outlined. Achievement
of this is assessed by a written report, requiring an appropriate
literature review of the concepts, and coherent explanation of
the observations and errors involved, as outlined in the
Supporting Information.
■
CONCLUSIONS
This experiment investigated the physical and chemical basis of
wettability of plant species in an undergraduate laboratory
setting. The concept of biomimetic research is introduced. The
four species of flowers chosen all resulted in being super-
hydrophobic. The high hysteresis values suggest the samples
collected were not self-cleaning. From this experiment the
group was able to demonstrate, and express understanding of,
the scientific principles that underpin the research area, and
they demonstrated acceptable levels of practical skill for final
year undergraduate students in capturing images using
microscopy techniques.
■
ASSOCIATED CONTENT
*
S Supporting Information
The Supporting Information is available on the ACS
Publications website at DOI: 10.1021/acs.jchemed.9b00324.
Lab notes for students (PDF, DOCX)
Lab notes for instructors (PDF, DOCX)
■ AUTHOR INFORMATION
Corresponding Author
*E-mail: chiara.neto@sydney.edu.au.
ORCID
Anthony Katselas: 0000-0003-0583-1367
Alice Motion: 0000-0002-5859-7888
Chiara Neto: 0000-0001-6058-0885
Notes
The authors declare no competing financial interest.
■
ACKNOWLEDGMENTS
The authors acknowledge funding from the Sydney Social
Sciences and Humanities Advanced Research Centre. The
authors thank Ahmed Owais for help in obtaining the SEM
micrographs (Figure 4).
■
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